Product details

Product details

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Immunogen

Prepared from whole rabbit serum produced by repeated immunizations with a synthetic peptide corresponding to aa 177-198 of mouse MDM2.

Specificity

This affinity-purified antibody is directed against the phosphorylated form of mouse MDM2 protein at the pS185 residue. The product was affinity purified from monospecific antiserum by immunoaffinity purification. Antiserum was first purified against the phosphorylated form of the immunizing peptide. The resultant affinity purified antibody was then cross-adsorbed against the non-phosphorylated form of the immunizing peptide. Reactivity occurs against Mouse MDM2 pS185 protein and the antibody is specific for the phosphorylated form of the protein. Reactivity with non-phosphorylated mouse MDM2 is minimal by ELISA and western blot. A BLAST analysis was used to suggest minimal cross reactivity with MDM2 homologues from other sources.Cellular Localization: Nuclear and cytoplasmic. Expressed predominantly in the nucleoplasm. Interaction with ARF (P14) results in the localization of both proteins to the nucleus. The nucleolar localization signals in both ARF and MDM2 may be necessary to allow efficient nucleolar localization of both proteins.

Purification

Immunoaffinity chromatography.

Target details

Target details

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Alternative Name

MDM2

Background

MDM2 is a nuclear phosphoprotein with an apparent molecular mass of 90 kD that forms a complex with the p53 tumor suppressor protein. Human MDM2 was identified as a homologous product of the 'murine double minute 2' gene (mdm2). The MDM2 gene enhances the tumorigenic potential of cells when it is overexpressed and encodes a putative transcription factor. Forming a tight complex with the p53 gene, the MDM2 oncogene can inhibit p53-mediated transactivation. MDM2 binds to p53 and amplification of MDM2 in sarcomas leads to escape from p53-regulated growth control. This mechanism of tumorigenesis parallels that for virusinduced tumors in which viral oncogene products bind to and functionally inactivate p53. Overexpression of the MDM2 oncogene was found in leukemias. Inactivation of tumor suppressor genes leads to deregulated cell proliferation and is a key factor in human tumorigenesis. MDM2 interacts physically and functionally with the retinoblastoma (RB) protein and can inhibit its growth regulatory capacity. Both RB and p53 can be subjected to negative regulation by the product of a single cellular protooncogene. The interference of binding to p53 prevents the interaction of MDM2 and its regulation of the transcriptional activity of p53 in vivo. Direct association of p53 with the cellular protein MDM2 results in ubiquitination and subsequent degradation of p53. MDM2-p53 complexes were preferentially found in S/G2M phases of the cell cycle. The MDM2 gene is alternatively spliced, producing 5 additional splice variant transcripts from the full length MDM2 gene. Four out of five of these alternatively spliced forms (MDM2a-MDMd) are missing substantial portions of the p53 binding domain and retain the acidic domain and the zinc-finger domains. The fifth and smallest transcript (MDM2e) retains the largest spliced region encoding the p53 binding domain, however, it lacks the nuclear localization signal, the acidic domain and zinc-finger domains. The alternatively spliced transcripts tend to be expressed in tumorigenic tissue, whereas the full-length MDM2 transcript is expressed in normal tissue.Synonyms: Double minute 2 protein, E3 ubiquitin-protein ligase Mdm2, Hdm2, Oncoprotein Mdm2, p53-binding protein Mdm2

Application Details

Application Details

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Application Notes

This affinity purified antibody has been tested for use in ELISA (1: 3,000 to 1: 12,000) andWestern blotting (1/500-1: 2,000). Western blotting shows specific banding forphosphorylated MDM 2 showing a band at approximately 102 kD corresponding tophosphorylated mouse MDM2. (See ref.5) for a discussion on expected molecular weights. Other applications not tested. Optimal dilutions are dependent on conditions and should be determined by the user.